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Cloning and analysis of the xylAB operon and characterization of xylose isomerase from Thermoanaerobacter ethanolicus

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Abstract

Three genes, xylA-like, xylA and xylB, were cloned and sequenced from the chromosome of Thermoanaerobacter ethanolicus JW200. xylA and xylB share an operon and encode xylose isomerase and xylulokinase, respectively. The xylA-like gene locates upstream of xylAB operon and encodes a hypothetical protein that lacks xylose isomerase activity. The xylose isomerase was expressed in Escherichia coli and purified by heat treatment and an ion-exchange chromatography. The enzyme had highest activity at 85°C and pH 7.0, and a half-life for 1 h at 85°C. The K m and V max values for xylose were 11 mM and 25 U/mg, respectively. The high level of expression, easy purification, and thermostability of the XylA from T. ethanolicus JW200 suggests industrial usefulness.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (no. 30770061).

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Authors and Affiliations

  1. Jiangsu Key Laboratory for Biodiversity and Bio-resources, College of Life Sciences, Nanjing Normal University, Nanjing, 210046, China

    Lijuan Fan, Yongchang Zhang, Wei Qu, Jingkai Wang & Weilan Shao

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  1. Lijuan Fan
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  2. Yongchang Zhang
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  3. Wei Qu
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  4. Jingkai Wang
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  5. Weilan Shao
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Correspondence to Weilan Shao.

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Fan, L., Zhang, Y., Qu, W. et al. Cloning and analysis of the xylAB operon and characterization of xylose isomerase from Thermoanaerobacter ethanolicus . Biotechnol Lett 33, 593–598 (2011). https://doi.org/10.1007/s10529-010-0463-x

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  • DOI: https://doi.org/10.1007/s10529-010-0463-x

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